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Please use this identifier to cite or link to this item: http://hdl.handle.net/1959.3/42357
- Title
- A method for prolonged imaging of motile lymphocytes
- Author(s)
- Day, Daniel; Pham, Kim; Ludford-Menting, Mandy J.; Oliaro, Jane; Izon, David J.; Russell, Sarah M.; Gu, Min
- Abstract
- With new imaging technologies and fluorescent probes, live imaging of cells in vitro has revolutionized many aspects of cell biology. A key goal now is to develop systems to optimize in vitro imaging, which do not compromise the physiological relevance of the study. We have developed a methodology that contains non-adherent cells within the field of view. 'Cell paddocks' are created by generating an array of microgrids using polydimethylsiloxane. Each microgrid is up to 250 x 250 μm2 with a height of 60 μm. Overlayed cells settle into the grids and the walls restrict their lateral movement, but a contiguous supply of medium between neighboring microgrids facilitates the exchange of cytokines and growth factors. This allows culture over at least 6 days with no impact upon viability and proliferation. Adaptations of the microgrids have enabled imaging and tracking of lymphocyte division through multiple generations of long-term interactions between T lymphocytes and dendritic cells, and of thymocyte-stromal cell interactions.
- Publication type
- Journal article
- Research centre
- Swinburne University of Technology. Faculty of Engineering and Industrial Sciences. Centre for Micro-Photonics
- Source
- Immunology and Cell Biology, Vol. 87, no. 2 (Feb 2009), pp.154-158
- Publication year
- 2009
- Keyword(s)
- Antigen presentation; Imaging; Lymphocytes; Microfabrication; T cells; Thymocytes
- Publisher
- Nature Publishing Group
- ISSN
- 0818-9641
- Publisher URL
- http://dx.doi.org/10.1038/icb.2008.79
- Copyright
- Copyright © 2009 Australasian Society for Immunology Inc. All rights reserved.
- Additional information
- This study was supported by the Australian Research Council, the National Health and Medical Research Council, the Human Frontier Science Program, and a Special Initiative Grant from Swinburne University of Technology.
- Peer reviewed
